CN109907304A - A kind of preparation method of Polysaccharide in Pleurotus eryngii - Google Patents
A kind of preparation method of Polysaccharide in Pleurotus eryngii Download PDFInfo
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- CN109907304A CN109907304A CN201910215140.3A CN201910215140A CN109907304A CN 109907304 A CN109907304 A CN 109907304A CN 201910215140 A CN201910215140 A CN 201910215140A CN 109907304 A CN109907304 A CN 109907304A
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Abstract
The invention discloses a kind of preparation methods of Polysaccharide in Pleurotus eryngii, belong to field of food and medicine.The present invention is a kind of method for combining proteinase combination zymolysis technique with pressurized hot water microwave leaching technology, improving Thick many candies content in Pleurotus eryngii water extract.Protein content in Pleurotus eryngii water extract can be reduced to 3% or less from 25% or so by this method, the low molecular weight impurities such as a large amount of monosaccharide, oligosaccharide, amino acid and small peptide are also removed simultaneously, Polysaccharide in Pleurotus eryngii purity is purified to 65% or more from the 20% of normal pressure boil water extraction method.Then, volatile alcohol is converted by the dextrose components in extracting solution using alcoholic fermentation, the purity of polysaccharide in Pleurotus eryngii extracting solution can be finally increased to 80% or more by drying processing removing alcohol.Compared with the technology of preparing of existing Polysaccharide in Pleurotus eryngii, Polyose extraction yield of the present invention improves 2~4 times, and whole process does not use organic solvent, and more preferably, purity is higher for the gas flavour of products obtained therefrom.
Description
Technical field
The present invention relates to a kind of preparation methods of Polysaccharide in Pleurotus eryngii, belong to field of food and medicine, can also be included into edible mushroom
Deep process technology field.
Background technique
Pleurotus eryngii (Pleurotus eryngiiQuel) the perverse celery of alias is picked up the ears, and is under the jurisdiction of Eumycota, Basidiomycetes, agaric
Mesh, Pleurotaceae, Pleurotus are that exploitation cultivation in recent years successfully integrates edible, medicinal, dietotherapy Rare edible fungus new product
Kind.Currently, domestic Pleurotus eryngii has realized factory culture, become a kind of common dish on common people's dining table.Planting almond abalone mushroom
Can be generated a large amount of by-product (such as misshapen mushroom, mushroom head) with process, yield up to 20% or more of total biomass, because
For mushroom head shapes are different and the inedible culture material of easy entrainment, it can not usually be sold and be abandoned as commodity.To the greatest extent
Pipe in this way, the Pleurotus eryngii mushroom head of enormous amount still can be used as Polysaccharide in Pleurotus eryngii prepare raw material, there is good development prospect.Separately
On the one hand, a large number of studies show that, Polysaccharide in Pleurotus eryngii have antitumor, anti-oxidant, antibacterial, reducing blood lipid, reducing blood lipid, norcholesterol,
The multiple biological activities such as antifatigue, anti-aging, value of exploiting and utilizing with higher.
Polysaccharide in Pleurotus eryngii generallys use the extraction of normal pressure burning water.However, the Pleurotus eryngii water extract obtained in this way
In polyoses content be usually less than 20%, protein impurities content is usually above 20%, non-polysaccharide carbohydrate (that is: monosaccharide and
Oligosaccharide) impurity content is up to 50% or more, and it is dense to heat the peculiar smell after boiling, subsequent products application is influenced, therefore need
Polysaccharide in Pleurotus eryngii product could be become by further purifying.
Non- polysaccharide component in Pleurotus eryngii boiling water water extract has: the protein of macromolecule and other small molecules (inorganic salts,
Monosaccharide, pigment, fat etc.).Wherein, trehalose impurity content can account for 50% of water extract or so, and protein content is 20% left
It is right.These small-molecule substances are dissolved in water mostly, can be removed by washing from residue, and most of insoluble proteins can also
To be first converted to small molecule amino acid and peptide with protease hydrolyzed, then removed from residue by washing.Due to variety classes
The digestion position of protease is different, so can not often obtain higher degree of hydrolysis using single protease, often adopts in practice
With multiple protein enzyme combined hydrolysis protein.
Since the polyoses content in normal pressure burning water extract is relatively low, to further increase Polysaccharide in Pleurotus eryngii content, need
The polysaccharide and protein in the ethanol precipitation solution of three times volume are recycled, the separation of small molecuies such as small molecule trehalose are gone out
It goes, then removes the protein in ethanol pellet with sevag method (that is: chloroform-n-butanol method), and then it is higher to obtain purity
Thick many candies product uses chromatographic column purification polysaccharide so as to a progress or is sold as polysaccharide health-care product.But this method is deposited
In obvious deficiency, this is primarily due to, and a large amount of volatile combustible organic solvents have been used in ethanol precipitation and Sevag method, molten
Relatively difficult, higher cost is recycled in agent, and has the risk of burning and explosion.
On the contrary, the present invention first uses enzymatic hydrolysis WATER-WASHING METHOD removal Polysaccharide in Pleurotus eryngii to extract protein and small point of solubility in raw material
Then it is lesser to be decomposed into molecular weight using Pintsch process effect by sub- impurity for insoluble macromolecular polysaccharide in Pleurotus eryngii residue
Soluble polysaccharide ingredient and monosaccharide (wherein: glucose accounts for 90% of monosaccharide component or more) and dissolve out, will using alcoholic fermentation
Monosaccharide in solution is converted into the alcohol of volatile, is enriched with polysaccharide component.Process of the present invention has without using solubility
Solvent, safety and environmental protection and cost is relatively low, equally available extract purity of polysaccharide with higher.Simultaneously as by egg
White matter digests and washing process, and the protein content and content of reducing sugar in Pleurotus eryngii extraction raw material substantially reduce, in conjunction with micro-
Acidic environment greatly reduces and generates Maillard reaction because of high-temp extracting, thus reduce the peculiar smell that is generated by high temperature and
Color burn phenomenon.By alcoholic fermentation, the gas flavour of extracting solution is also greatly improved.
Summary of the invention
It is an object of the invention to be directed to the deficiency of existing normal pressure extracting in boiling water Polysaccharide in Pleurotus eryngii technology, it is to provide for one kind
Improved Polysaccharide in Pleurotus eryngii preparation method.Compared with existing normal pressure boil water extraction method combination sevag deproteinized technology of preparing, this
The polysaccharide preparation process of invention does not use organic solvent, and extraction efficiency is higher, and product purity equally with higher.
To achieve the above object, the present invention uses following technological means:
A kind of preparation method of Polysaccharide in Pleurotus eryngii hydrolyzes Pleurotus eryngii first with two or more proteinase combinations, will be in Pleurotus eryngii
Insoluble protein be hydrolyzed into soluble peptide or amino acid after be filtered to remove；Then gained is digested into the pure washing of residue
For several times, protein zymolyte, trehalose and other soluble non-polysaccharide impurity are washed away；Then rinsing residue is placed in reaction under high pressure
It in kettle, is extracted using pressurized hot water, after extraction, by leaching liquor filtering, collects filtrate high pressure sterilization, inoculation distillery yeast hair
Ferment first converts volatile alcohol for the monosaccharide in extracting solution, then by drying and removing alcohol, is enriched with polysaccharide, most
Obtaining eventually has higher degree and the light Pleurotus eryngii Thick many candies powder of smell.
Specifically includes the following steps:
(1) proteinase combination digests
Fresh Pleurotus eryngii or Pleurotus eryngii processing byproduct are dried, clays into power, water dispersion is added with 1:10~1:40 solid-to-liquid ratio, is made
The pH value of dispersion liquid is adjusted to the appropriate amount of the first protease hydrolyzed with 1 mol/L NaOH or HCl solution, is added the
A kind of protease, is placed in water-bath, is digested with suitable temperature, 1 mol/L NaOH solution is during which constantly added dropwise, to protect
It is constant to hold solution ph, after enzymatic hydrolysis, heating, which is boiled, makes albumen enzyme-deactivating；It is molten using 1 mol/L NaOH or HCl after cooling
Liquid adjusts the pH value of dispersion liquid to the appropriate amount of second of protease hydrolyzed, and second of protease hydrolyzed, enzymatic hydrolysis knot is added
Shu Hou, heating, which is boiled, makes albumen enzyme-deactivating；Solid residue is collected after filtering, discards filtrate；
(2) it washes
The deionized water of 5~10 times of weight of enzymatic hydrolysis residue obtained by step (1) is washed 3~5 times, after filter-cloth filtering, is received
Collect solid residue, discards filtrate；
(3) pressurized hot water extracts
It is placed in step (2) is residue obtained in autoclave, the deionized water of 20 times of weight is added, dispersion liquid is made, uses 1
Mol/L HCl solution adjusts the pH value of dispersion liquid to subacidity, and pressurization extracts, and cooled and filtered obtains filtrate, discards residual
Slag；
(4) alcoholic fermentation
By clear liquid rotary evaporation obtained by step (3), being concentrated into solid content is 6~10wt%, uses 1 mol/L HCl solution
Adjusting solution ph is 4.5, is inoculated with saccharomyces cerevisiae under aseptic condition, is placed in 32 DEG C of constant incubators 48 h of fermentation, is then centrifuged
Separation discards ferment mud, collects supernatant；
(5) dry
The pH value of the fermented supernatant fluid of step (4) is adjusted to 7 with 1 mol/L NaOH solution, is then dried, that is, obtains
Pleurotus eryngii Thick many candies powder.
Pleurotus eryngii powder is 100 ~ 200 mesh in step (1).
Protease as described in step (1) is alkali protease, compound fertilizer production, neutral proteinase, Papain
One of enzyme, trypsase, bromelain, ficin, acid protease or pepsin.
Extracting technology parameter in step (3) are as follows: initial pH 4~6, extraction temperature are 155~165 DEG C, extraction time 1~
60 min。
Inoculation saccharomyces cerevisiae concentration is 10 in step (4)9 ~1010Cfu/mL, inoculum concentration are 10%~20%, fermentation temperature
It is 28~32 DEG C, fermentation time is 48~72 h.
The beneficial effects of the present invention are:
1, after applying the present invention, compared with the technology of preparing of existing Polysaccharide in Pleurotus eryngii, Polyose extraction yield of the present invention improves 2~4
Times；
2, a large amount of strong acid is generated without using organic solvent, not；Have the advantages that it is economical, nontoxic, will not combustion explosion；
3, under the conditions of pressurized high-temperature, since the solubility of polysaccharide in water is greatly improved, and pressurized high-temperature is extracted
Liquid during being cooled to room temperature, middle and high concentration, the polysaccharide in hypersaturated state can still keep dissolved state, therefore make
Polysaccharide concentration may be up to 2% or more in standby obtained pressurized high-temperature extracting solution, be much higher than normal pressure extracting in boiling water liquid 0.3%；Therefore,
Aqueous solution volume needed for extracting same quality polysaccharide greatly reduces, and this greatly reduces the difficulty of subsequent polysaccharide dehydrating operations
And cost, it is easier to realize industrialization；
4, it after using polysaccharide technology is extracted after first deproteinized of the present invention, in conjunction with adjustment solution acidic environment, is produced by high temperature
Raw Maillard reaction is inhibited；Pleurotus eryngii Thick many candies combinations color and lustre is shallower, peculiar smell is lighter, has and is more widely applied model
It encloses.
5, by alcoholic fermentation, the monosaccharide in high pressure extraction liquid is eliminated, not only obtains the purity of Polysaccharide in Pleurotus eryngii product
To further increasing, and product gas flavour is improved, and the peculiar smell generated by " mushroom " taste is inhibited.
Specific embodiment
Below in conjunction with specific embodiment, the present invention will be further described, but the present invention is not limited only to these embodiments.
Embodiment 1
(1) proteinase combination digests
Fresh Pleurotus eryngii or Pleurotus eryngii processing byproduct are dried, clays into power, sieves with 100 mesh sieve, moisture is added with 1:100 solid-to-liquid ratio
Dissipate, using the pH value of 1 mol/L NaOH solution adjusting dispersion liquid to pH 8, with 2%(w:w) ratio addition alkali protease,
It is placed in water-bath, is digested 2 hours with 55 DEG C, 1 mol/L NaOH solution is added dropwise, constantly during which to keep solution ph not
Become, after enzymatic hydrolysis, heating, which is boiled, makes albumen enzyme-deactivating.After cooling, the soda acid of dispersion liquid is adjusted using 1 mol/L HCl solution
Degree is to pH 6, with 1%(w:w) ratio addition compound fertilizer production enzymatic hydrolysis, it is digested 2 hours with 45 DEG C, after enzymatic hydrolysis, heating
Boiling makes albumen enzyme-deactivating.Solid residue is collected after filtering, discards filtrate；
(2) it washes
The deionized water of 5 times of weight of enzymatic hydrolysis residue obtained by step (1) is washed 3 times, after filter-cloth filtering, it is residual to collect solid
Slag discards filtrate；
(3) pressurized hot water extracts
It is placed in step (2) is residue obtained in autoclave, the deionized water of 20 times of weight is added, dispersion liquid is made, uses 1
Mol/L HCl solution adjusts the pH value of dispersion liquid to pH 6, is extracted with 155 DEG C of pressurized high-temperatures, cooled and filtered is filtered
Liquid discards residue；
(4) alcoholic fermentation
By clear liquid rotary evaporation obtained by step (3), being concentrated into solid content is 6wt%, is adjusted using 1 mol/L HCl solution
Solution ph is 4.5, is inoculated with saccharomyces cerevisiae under aseptic condition, is placed in 32 DEG C of constant incubators and ferments 48 h, then 5000rpm from
The heart separates 5min, discards ferment mud, collects supernatant.
(5) dry
The pressurized hot water leaching liquor pH value of step (4) is adjusted to 7 with 1 mol/L NaOH solution, at freeze-drying
Reason, i.e. acquisition Pleurotus eryngii Thick many candies powder.
After this method, the polyoses content in Pleurotus eryngii water extract can be increased to from the 20% of normal pressure boil water extraction method
80%, the extraction yield of polysaccharide is increased to 35%(from 8% and is counted using drying Pleurotus eryngii as raw material), color is shallower, and peculiar smell is lighter.
Embodiment 2
(1) proteinase combination digests
Fresh Pleurotus eryngii or Pleurotus eryngii processing byproduct are dried, clayed into power, 200 meshes is crossed, moisture is added with 1:20 solid-to-liquid ratio
Dissipate, using the pH value of 1 mol/L NaOH solution adjusting dispersion liquid to pH 7, with 2%(w:w) ratio, neutral proteinase is added,
It is placed in water-bath, is digested 2 hours with 50 DEG C, 1 mol/L NaOH solution is added dropwise, constantly during which to keep solution ph not
Become, after enzymatic hydrolysis, heating, which is boiled, makes albumen enzyme-deactivating.After cooling, the soda acid of dispersion liquid is adjusted using 1 mol/L HCl solution
Spend to pH 6, then with 1.5 %(w:w) ratio, papain enzymolysis is added, 50 DEG C digest 2 hours, after enzymatic hydrolysis, add
Heat, which is boiled, makes albumen enzyme-deactivating.Solid residue is collected after filtering, discards filtrate；
(2) it washes
The deionized water of 5~10 times of weight of enzymatic hydrolysis residue obtained by step (1) is washed 3~5 times, after filter-cloth filtering, is received
Collect solid residue, discards filtrate；
(3) pressurized hot water extracts
It is placed in step (2) is residue obtained in autoclave, the deionized water of 20 times of weight is added, uses 1 mol/L HCl
Solution adjusts the pH value of dispersion liquid to pH 5, is extracted with 165 DEG C of pressurized high-temperatures, cooled and filtered obtains filtrate, discards residual
Slag；
(4) alcoholic fermentation
By clear liquid rotary evaporation obtained by step (3), being concentrated into solid content is 8wt%, is adjusted using 1 mol/L HCl solution
Solution ph is 4.5, is inoculated with saccharomyces cerevisiae under aseptic condition, is placed in 32 DEG C of constant incubators 36 h of fermentation, then 5000 rpm
10 min are centrifugated, ferment mud is discarded, collect supernatant.
(5) dry
The pressurized hot water leaching liquor pH value of step (4) is adjusted to 7 with 1 mol/L NaOH solution, at spray drying
Reason, i.e. acquisition Pleurotus eryngii Thick many candies powder.
After this method, the polyoses content in Pleurotus eryngii water extract can be increased to from the 20% of normal pressure boil water extraction method
65%, protein impurities content is reduced to 3.5% from 20%, and the yield for extracting polysaccharide is increased to 38%(from 6% with drying Pleurotus eryngii and is
Raw material meter), color is shallower, and peculiar smell is lighter.
Claims (6)
1. a kind of preparation method of Polysaccharide in Pleurotus eryngii, it is characterised in that: hydrolyze apricot Bao first with two or more proteinase combinations
Mushroom is filtered to remove after the insoluble protein in Pleurotus eryngii to be hydrolyzed into soluble peptide or amino acid；Then gained is digested
Residue for several times, washes away protein zymolyte, trehalose and other soluble non-polysaccharide impurity with pure washing；Then it will wash residual
Slag is placed in autoclave, is extracted using pressurized hot water, after extraction, by leaching liquor filtering, collect filtrate high pressure sterilization,
It is inoculated with distillery yeast fermentation, first converts volatile alcohol for the monosaccharide in extracting solution, then by drying and removing alcohol, make more
Sugar is enriched with, and final obtain has higher degree and the light Pleurotus eryngii Thick many candies powder of smell.
2. the preparation method of Polysaccharide in Pleurotus eryngii according to claim 1, it is characterised in that: specifically includes the following steps:
(1) proteinase combination digests
Fresh Pleurotus eryngii or Pleurotus eryngii processing byproduct are dried, clays into power, water dispersion is added with 1:10~1:40 solid-to-liquid ratio, is made
The pH value of dispersion liquid is adjusted to the appropriate amount of the first protease hydrolyzed with 1 mol/L NaOH or HCl solution, is added the
A kind of protease, is placed in water-bath, is digested with suitable temperature, 1 mol/L NaOH solution is during which constantly added dropwise, to protect
It is constant to hold solution ph, after enzymatic hydrolysis, heating, which is boiled, makes albumen enzyme-deactivating；It is molten using 1 mol/L NaOH or HCl after cooling
Liquid adjusts the pH value of dispersion liquid to the appropriate amount of second of protease hydrolyzed, and second of protease hydrolyzed, enzymatic hydrolysis knot is added
Shu Hou, heating, which is boiled, makes albumen enzyme-deactivating；Solid residue is collected after filtering, discards filtrate；
(2) it washes
The deionized water of 5~10 times of weight of enzymatic hydrolysis residue obtained by step (1) is washed 3~5 times, after filter-cloth filtering, is received
Collect solid residue, discards filtrate；
(3) pressurized hot water extracts
It is placed in step (2) is residue obtained in autoclave, the deionized water of 20 times of weight is added, dispersion liquid is made, uses 1
Mol/L HCl solution adjusts the pH value of dispersion liquid to subacidity, and pressurization extracts, and cooled and filtered obtains filtrate, discards residual
Slag；
(4) alcoholic fermentation
By clear liquid rotary evaporation obtained by step (3), being concentrated into solid content is 6~10wt%, uses 1 mol/L HCl solution
Adjusting solution ph is 4.5, is inoculated with saccharomyces cerevisiae under aseptic condition, is placed in 32 DEG C of constant incubators 48 h of fermentation, is then centrifuged
Separation discards ferment mud, collects supernatant；
(5) dry
The pH value of the fermented supernatant fluid of step (4) is adjusted to 7 with 1 mol/L NaOH solution, is then dried, that is, obtains
Pleurotus eryngii Thick many candies powder.
3. the preparation method of Polysaccharide in Pleurotus eryngii according to claim 2, it is characterised in that: Pleurotus eryngii powder in step (1)
For 100 ~ 200 mesh.
4. the preparation method of Polysaccharide in Pleurotus eryngii according to claim 2, it is characterised in that: albumen as described in step (1)
Enzyme be alkali protease, compound fertilizer production, neutral proteinase, papain, trypsase, bromelain, without flower
One of fruit protease, acid protease or pepsin.
5. the preparation method of Polysaccharide in Pleurotus eryngii according to claim 2, it is characterised in that: extracting technology is joined in step (3)
Number are as follows: initial pH 4~6, extraction temperature are 155~165 DEG C, 1~60 min of extraction time.
6. the preparation method of Polysaccharide in Pleurotus eryngii according to claim 2, it is characterised in that: inoculation wine brewing ferment in step (4)
Female concentration is 109 ~1010Cfu/mL, inoculum concentration are 10%~20%, and fermentation temperature is 28~32 DEG C, and fermentation time is 48~72
h。
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Application publication date: 20190621 |